KR20160058845A - Novel peptides and analogs for use in the treatment of oral mucositis - Google Patents

Abstract

Preclinical data obtained in a model of chemotherapy-induced mucositis, radiation-induced mucositis, neutropenia infection and colitis indicate that oral mucositis is a potent indication for a congenital defense regulator (IDR) peptide. The clinical efficacy results obtained with IDR in the mouse and hamster models of mucositis indicate that every 3 days of dosing should be able to cover the mucositis "window " at doses of 7 to 14, depending on the duration of chemotherapy or radiation exposure. IDR also demonstrated efficacy in mouse models of chemotherapy-induced oral and gastrointestinal mucositis, consistent with the response of innate immune responses to chemotherapy and / or radiation injury. IDR is also effective in reducing bacterial burden in the presence or absence of antibiotic therapy in a variety of infectious and infectious models and enhances survival.

Description

[0001] The present invention relates to novel peptides and analogs for use in the treatment of oral mucositis. ≪ RTI ID = 0.0 > [0002] <

Related application

This application claims priority from U.S. Provisional Application No. 61 / 877,767 filed on September 13, 2013, the contents of which are incorporated herein by reference.

Introduction

Congenital immune system

Congenital immune responses are an evolutionarily conserved protection system associated with barriers between the tissue and the external environment, such as skin, mucous membranes and airways. It is often associated with inflammatory responses and is a major contributor to the activation of adaptive immunity, when prompt recognition and eradication of pathogen infiltration as well as responses to cellular damage is provided. Congenital defenses are triggered by the binding of pathogens and / or damage-associated molecules (PAMP or DAMP) to pattern-cognitive receptors containing toll-like receptors (TLRs). Pattern cognitive receptors are found on many cell types and on many cell types and are distributed throughout the body in both the circulatory and tissue resident compartments and act to provide an initial "dangerous" Release of non-specific antimicrobial molecules, cytokines, chemokines, and host defense proteins and peptides, as well as mobilization of immune cells (neutrophils, macrophages, monocytes) (Janeway 2002; Beutler 2003; Beutler 2004; Athman 2004; Tosi 2005 ; Doyle 2006; Foster 2007; Matzinger 2002). In addition, the innate immune system is directly involved in the development of tolerance to symbiotic microbial guns in the gastrointestinal tract, and gastrointestinal recovery and immune defense (Santaolalla, 2011; Molloy, 2012).

Mucositis

Mucositis is a clinical term for damage done to the mucosal layer by chemotherapy. It can occur in any mucosal layer area, but is most commonly associated with the mouth, and then with the small intestine. While many mucositis grades are used clinically, the two most commonly used grading systems are NCI and WHO grades.

Mechanisms of mucositis have been extensively studied and have recently been implicated in the interaction of chemotherapy and / or radiotherapy with congenital defense systems (Sonis 2010). Bacterial infections of ulcerative lesions are now regarded as secondary consequences of regulatory impaired local inflammation triggered by therapy-induced cell death rather than as a major cause of lesions. Mucositis affects 500,000 humans annually in the US and occurs in 40% of patients treated with chemotherapy (Sonis 2010, Curr. Op.). Mucositis most commonly occurs in patients with head and neck cancer treated with radiotherapy (> 80% of severe mucosal lesions) (Elting et al. 2008). Mucositis is common in patients treated with high-dose chemotherapy and stem cell transplantation (SCT) (40-100% incidence), where the onset and severity of mucositis is characterized by the nature of the conditioning regimen used for bone marrow injury It is highly dependent (Murphy 2007). Among well-established chemotherapeutic drugs, 5-FU and irinotecan are particularly well known for causing mucositis, but they also occur with newer agents such as mTOR inhibitors and kinase inhibitors (Mateus et al. 2009; Sankhala et al 2009). Mucositis can be severely compromised and can lead to the need for infection, sepsis, parenteral nutrition and narcotic analgesics. Intestinal damage causes severe diarrhea. These symptoms can limit the dose and duration of the cancer treatment, leading to a second-line treatment that includes reduced survival. Direct and indirect results of mucositis were estimated to add ~ $ 18K per patient to cancer treatment costs (Nonzee et al. 2008). Mucositis occurs 3-12 weeks after radiation onset or 3-12 days after chemotherapy initiation, and disappears after 2-3 weeks unless further chemotherapy or radiation therapy is performed.

RIVPA (SEQ ID NO. 5) is an IDR (Innate Defense Regulator), a new class of short synthetic peptides with a novel mechanism. IDR, designed to mimic one of the recently discovered functions of natural mucosal defense peptides, has no direct antibiotic activity, but can modulate the host response to increase survival after infection with a wide range of bacterial gram-negative and gram-positive pathogens But accelerates the resolution of tissue damage after exposure to a variety of agents, including bacterial pathogens, trauma and chemotherapy or radiation therapy.

Based on preclinical data obtained in models of chemotherapy-induced mucositis, radiation-induced mucositis, neutrophil infections and colitis, oral mucositis is a potent symptom in RIVPA (SEQ ID NO. 5) and other IDR peptides. The IV dosage form of RIVPA (SEQ ID NO. 5) is well suited for mucositis indications since the drug will be provided soon after chemotherapy input or radiation. The clinical efficacy results obtained with RIVPA (SEQ ID NO. 5) in a mouse and hamster model of mucositis can cover mucositis "windows " by administration every 3 days, depending on chemotherapy or radiation exposure, 7-14 times .

In relation to breast cancer, ~ 20% of patients treated with ACT therapy suffer from ulcerative mucositis during their first chemotherapy, and ~ 70% of those patients are treated twice with their treatment And have ulcerative mucositis (Sonis 2010). This represents a beneficial "high risk" patient population from treatment with RIVPA (SEQ ID NO. 5). There is no systemic agent currently available for the improvement of mucositis in these communities.

Patients treated with high-dose chemotherapy and SCT for the treatment of blood cancer are highly immunosuppressed populations at risk of infection. In such treatment, a "pretreatment" which is a high dose of chemotherapy (sometimes combined with radiation) is used to kill a high percentage of cancer cells. These levels of treatment will result in lethal bone marrow suppression unless stem cells (from bone marrow or blood) are administered later to allow reconstitution of blood cells. Autotransplantation uses the patient's own stem cells for this purpose, while allografts use cells from matched healthy donors. Autologous transplantation is most often used to treat multiple myeloma (MM) and non-Hodgkin's lymphoma (NHL). Allografts are typically used to treat leukemia, such as AML.

Regarding SCT, until recently, a variety of pre-treatment chemotherapy treatments have resulted in a relatively high rate of oral mucositis (40-100%), and in most US centers these patients are admitted. Oral mucositis associated with radiation and / or chemoradiotherapy for head and neck cancer is a major problem with 85% of subjects suffering from some mucositis - 42% is grade 3 or 4.

Acute radiation syndrome

Acute Radiation Syndrome (ARS) is a serious disease that occurs when the entire body (or most of it) receives high doses of radiation, typically over a short period of time. Many survivors of the atomic bombs of Hiroshima and Nagasaki in the 1940s and many firefighters who first responded to the Chernobyl nuclear accident in 1986 became ill with ARS (CDC 2013).

Individuals exposed to radiation will only get ARS if:

• radiation doses are high (doses from medical procedures such as chest X-rays are so low that they can not cause ARS)

Radiation penetrates (i.e., can reach the internal organs)

The entire human body or a majority of it accepts doses,

Radiation was generally received within a few minutes in a short time.

Radiation causes dose-proportional injury to mammalian cells and tissues. At low doses, injury can be limited to long-term effects, such as point mutations in somatic cells and / or germline DNA that may be associated with increased risk of cancer or birth defects. At moderate doses, radiation causes chromosomal anomalies, such as breakdown and dislocation, which again increases the risk of cancer and birth defects and, if severe enough, will result in rapid cell differentiation within the time of exposure. At very high doses, radiation can denature proteins and cause most immediate lethality of cells and tissues. Tissues with rapidly differentiating cells, most commonly affected by moderate dose radiation, include bone marrow, gastrointestinal tract, and testes. Exposure to radiation can be used to treat and prevent gastrointestinal toxicities such as, for example, diarrhea and more chronic effects, such as acute effects including skin rash and burn, anemia, decreased leukocyte count and thrombocytopenia, Development and birth defects.

The first symptoms of ARS are typically nausea, vomiting, and diarrhea. These symptoms will begin within minutes to a few days after exposure, last for several minutes to several days, and may disappear. After that, humans generally appear and feel healthy for a short period of time, and then he or she is sick again and falls into anorexia, fatigue, fever, nausea, vomiting, diarrhea and possibly even seizures and coma. Such severe disease stages may last from several hours to several months.

Humans with ARS typically also have some skin damage. Such damage can begin to appear within hours after exposure and can include skin swelling, itching, redness and hair loss. As with other symptoms, the skin can heal for a short time and then cause swelling, itching and redness after a few days or weeks. Complete healing of the skin can take months to years depending on the dose of radiation the human skin has received.

The gastrointestinal symptoms of ARS are referred to as gastrointestinal acute radiation syndrome or GI-ARS. GI-ARS consists of diarrhea, dehydration, enterobacterial infection, and, in severe cases, septic shock and lethal (Potten 1990). After radiation exposure, GI-ARS is thought to cause direct damage to the basal stem cells of the crypts of Lieberkuhn, leading to mitotic arrest and death through apoptotic mechanisms (Potten 1997a, Potten 1997b). The integrity of the gastric mucosa depends on the rapid proliferation of pluripotent stem cell pools at the bottom of the crypt (Brittan 2002, Gordon 1994, Potten 1997b). Thus, stem cell lethality is considered to be a crucial factor in this process, as the surviving intestinal stem cells appear to be sufficient for the reconstitution of the vagina and the villi (Potten 1990). Regeneration of intestinal epithelial barrier is dependent on active stem cell compartments similar to hematopoietic cells. Long-term precursor clonogenic cells are particularly susceptible to ionizing radiation exposure, and as radiation doses increase, and -chromosome cloning cells can not produce enough cells to reproduce the villi. This leads to cloudiness and diminishment of villus height and ultimately to functional impairment leading to decreased nutrient absorption and barrier function, fluid and electrolyte loss, and bacterial disruption through intestinal barriers (Monti 2005, Zhao 2009). Above 8 gray (Gy), dose-dependent stem cell lethality leads to a reduction in regeneration until the regeneration level is insufficient to rescue the GI mucosa. In mouse studies, progressive peeling of the epithelium leads to lethality from GI syndrome 6 to 7 days post radiation. When mitotic activity is resumed, a sudden depletion of < RTI ID = 0.0 > and, < / RTI > In the lower-dose range (8-13 Gy), the surviving proliferating cells regenerate the system and lead to complete regeneration of the damaged mucosa. At doses exceeding 14 Gy, a large amount of proliferative cell loss results in animal death from the collapse of the viral system, mucosal peeling and gastrointestinal syndrome (Paris 2001; Potten 1990; Withers 1971; Withers 1969).

The intestinal stem cell compartment is not the only compartment sensitive to ionizing radiation. Another critical factor that involves the response of the GI tract to major physical attacks is the hypersalivation of the intestine. Ongoing persistent hypopigmentation is an important stimulus event in the development of systemic inflammatory response syndrome and multiple organ failure (MOF) (Moore, 1999). Increased intestinal vascular permeability with capillary leakage was observed early in the study (Cockerham 1984; Eddy 1968, Willoughby 1960). Additional irradiation-posterior deformations include late bleeding patterns following mid-dilation and torsion of the small artery, reduction of blood vessel count and / or length (Eddy 1968). There is a continuing debate about whether the primary challenge is the result of lethal epithelial stem cell death or endothelial cell death (Kirsch 2010). Regardless of the original challenge, it is clear that the challenge causes a complex injury response involving intestinal epithelial cells, lethal endothelial cells, and proximal humoral immune response (Williams 2010).

Therapeutic Methods For example, hematopoietic growth factors, i.e., granulocyte- and / or granulocyte-macrophage colony stimulating factors (G-CSF and G / M-CSF) and erythropoietin (EPO), and hematopoietic stem cell / It is possible to weaken the mortality rate from dysfunction.

Human survival opportunities with ARS are reduced with increasing radiation doses. The causes of deaths within 15 days of exposure to radiation are generally injuries to the GI tract, whereas deaths after 15 days are generally the result of bone marrow injury. For survivors, the recovery process can last from weeks to two years (CDC 2013).

Currently, there is no approval for the treatment of acute radiation syndrome, so development of radiation emollients is urgently required. RIVPA (SEQ ID NO. 5) has the potential to reduce acute mortality in ARS, enabling a management effort and helping to restore skin damage.

infection

A variety of microorganisms, including viruses, bacteria, fungi and parasites, can cause disease. Microbial cells are distinguishable from animal and plant cells, which can not survive alone in nature and exist only as part of multicellular organisms. The cells of the microorganism may be pathogenic or non-pathogenic, depending in part on the microorganism and the host condition. For example, in immunocompromised hosts, generally harmless bacteria can be pathogens. Entry into host cells is important for the survival of bacterial pathogens that replicate in the intracellular milieu. For individuals replicating at extracellular locations, the importance of bacterial entry into host cells is not well understood.

Drug tolerance is an obstacle to ongoing efforts to combat infection. For example, penicillin is effective in treating Staphylococcus aureus , but is effective only until the bacteria have tolerance. Over the second half of the 20th century, new antibiotics such as vancomycin and methicillin have been developed; These antibiotics successfully treated S. aureus infection. However, in the 1970s S.Aureus Methicillin-resistant strains have evolved and have since become widespread in hospitals worldwide. More recently, S. aureus Vancomycin-resistant species emerged.

With the increasing threat of resistance to antimicrobial drugs and the emergence of new infectious diseases, there is a continuing need for novel therapeutic compounds. Therapeutic agents that work in the host but not in the pathogen are preferred because they do not promote the resistance of the pathogen. In particular, drugs acting on the host through the innate immune system provide a source of potential therapeutic agents. There is evidence that congenital immune responses are the means to control most infections and also contribute to inflammatory responses. Inflammation-induced inflammatory responses are known to be a key component of pathogenesis. The ability to control inflammation, while increasing host resistance to infection, would be highly beneficial in the ongoing fight against infection, including infection, caused by resistant bacteria.

IDR  And congenital immune system

Congenital defense regulators (IDRs) interact with intracellular signaling events and regulate congenital defense responses. While many early work with IDR focused on their role in fighting infections while controlling inflammation, recent results in animal models of chemotherapy- or radiation-induced mucositis and wound healing suggest that IDR is a pathogen- Suggesting that it may be beneficial during the reaction to the derivatizing agent. IDR treats and prevents infection by selectively modifying the body's innate defense response without triggering the associated inflammatory response when they are activated by PAMP or DAMP (Matzinger 2002). The same mechanism is the basis for the positive effects observed in mucositis and wound healing models, where DAMP or subsequent signaling is affected. RIVPA (SEQ ID NO. 5) has been shown to be safe in humans and in animal models of split radiation-induced and chemotherapy-induced oral mucositis, chemotherapy induced damage models for gastrointestinal tract and localization in immunologically and immunocompromised hosts. And whole body Gram-positive and Gram-negative infection models.

FIG. RIVPA (SEQ ID NO. 5) reduces the severity of oral mucositis in the split-irradiation model.
FIG. RIVPA (SEQ ID NO. 5) reduces the severity of severe oral mucositis in divided-dose models using optimal dosing regimens.
FIG. RIVPA (SEQ ID NO. 5) decreases severity of DSS-induced colitis measured by endoscopy (A) on days 7, 14 and 21 and histopathology (B, C, D)
FIG. RIVPA (SEQ ID NO. 5) reduces the duration of severe oral mucositis (A), severity of colitis (B) and weight loss (C) in the chemotherapy model (Study 1).
Figure 5 . RIVPA (SEQ ID NO. 5) reduces the duration of severe oral mucositis (A), severity of colitis (B) and weight loss (C) in the chemotherapy model (Study 2).
FIG. RIVPA (SEQ ID NO. 5) decreases the duration of severe oral mucositis (A), severity of colitis (B) and weight loss (C) in a dose-response chemotherapeutic model.
FIG. Combination of RIVPA (SEQ ID NO. 5) with vancomycin therapy in an MRSA IP infectious model.
Figure 8 . RIVPA (SEQ ID NO. 5) activity in neutropenic mice of a thigh abscess MRSA infection model.
Figure 9 . The dose response of RIVPA (SEQ ID NO. 5) in the MRSA bacteraemia model of immunologically competent mice.
10. The dose response of RIVPA (SEQ ID NO. 5) in MRSA bacteremia models of T-cell deficient mice.
11. Therapeutic RIVPA (SEQ ID NO. 5) efficacy in S. aureus acute peritoneal infection model.
12. RIVPA (SEQ ID NO. 5) activity in neutropenic mice of the thigh abscess S. aureus infection model.
Fig. RIVPA (SEQ ID NO. 5) Efficacy in a Klebsiella Peritoneal Infection Model with High (A) and Low (B) Bacterial Infections. * In (A), the absence of the bar indicates that all mice died (0% survival).
14. RIVPA (SEQ ID NO. 5) improves the resolution of tissue damage in locally MRSA-infected skin. 48h Bacterial Burden (A), 96h Scatter plot (B), Blinded Photographic Scoring, 48h (C), Blind Photo Scoring, 96h (D). * The bars in (C) and (D) indicate that all mice have a grade of 0, yielding a standard error (SEM) of the mean of 0 ± 0 and an average.
15. A lack of RIVPA (SEQ ID NO. 5) for the recovery of circulating blood cell leukocytes (A) or neutrophils (B) after the introduction of leukopenia in CD-1 mice.
Fig. R (tBg) V1KR (tBg) V2 (SEQ ID NO. 91) decreases the severity of oral mucositis in the chemotherapeutic model.
Fig. RIV (mp2) A-NH2 (SEQ ID NO. 92) reduces the severity of oral mucositis in a chemotherapeutic model.
Fig. R (tBg) V1KR (tBg) V2 (SEQ ID NO: 91) increases survival in MRSA bacteraemia models.
Fig. RIV (mp2) A-NH2 (SEQ ID NO. 92) increases survival in MRSA bacteremia models.

DETAILED DESCRIPTION OF THE INVENTION

It is an object of the present invention to provide a discrete peptide consisting of the amino acid sequence of R (tBg) V1KR (tBg) V2 wherein tBg = tert-butylglycine and further R (tBg) Amide bond.

이다.It is an object of the present invention to provide a separate peptide consisting of the amino acid sequence of RIV (mp2) A-NH2 wherein mp2 = 4-amino-1-methyl-1H-pyrrole-

to be.

A still further object of the present invention is a method of treating oral mucositis in a subject exposed to radiation or a chemotherapeutic agent in a damaging amount,

a) a peptide comprising the amino acid sequence of Table 1; or

b) SEQ ID NOs: 5,7,10,14,17,18,22,23,24,27,31,34,35,63,64,66-69,72,76,77,90, 91 and 92, or a pharmaceutical salt, ester or amide thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

It is an object of the present invention to provide a method of treating oral mucositis in a subject exposed to a radiation or chemotherapeutic agent,

a) a peptide comprising the amino acid sequence of amino acids of more than _{7, X 1 X 2 X 3 P} (SEQ ID NO: 56) ( herein, X1 is R a; X2 is I or V, where X2 is X3 is I or V, wherein X3 can be N-methylated; P is a proline or proline analogue; SEQ ID NO: 56 is the first four amino acids of the N-terminus of the peptide ) Or a pharmaceutical salt, ester or amide thereof and a pharmaceutically acceptable carrier, diluent or excipient; or

b) a polynucleotide having the nucleotide sequence of SEQ ID NO: 5, 7, 10, 14, 17, 18, 22, 23, 24, 27, 28, 31, 34, 35, 63, 64, 66-69, 72, 92, or a pharmaceutical salt, ester or amide thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

A further object of the present invention is a method of treating oral mucositis in a subject exposed to a radiation or chemotherapeutic agent in a quantity wherein the peptide is selected from the group consisting of SEQ ID NO: 5 or a pharmaceutical salt, ester or amide thereof, Carrier, diluent, or excipient.

It is yet another object of the present invention to provide a method for treating oral mucositis in a subject exposed to a radiation dose or chemotherapeutic agent, wherein the peptide is administered orally, parenterally, transdermally, or intranasally.

It is yet another object of the present invention to provide a method of treating oral mucositis in a subject exposed to a radiation or chemotherapeutic agent, wherein the effective amount of the peptide administered to the subject is at least 1 mg / kg. In a preferred embodiment, the effective amount of the peptide administered to the subject is from about 1.5 mg / kg to about 6 mg / kg.

It is yet another object of the present invention to provide a method of treating oral mucositis in a subject exposed to a radiation or chemotherapeutic agent in a wound, the method comprising administering the peptide to a subject every three days during the administration of radiation or chemotherapeutic agents will be.

It is yet another object of the present invention to provide a method of treating oral mucositis in a subject exposed to a radiation or chemotherapeutic agent in an amount wherein the peptide is combined with a locally active corticosteroid or a metabolite thereof in an oral dosage form Administered to a subject, wherein the oral dosage form is effective for topical or local treatment of the gastrointestinal tract and oral cavity of the subject, and the subject further provides a method of targeting a subject that exhibits inflammatory symptoms due to tissue damage resulting from radiation or chemotherapy treatment . Representative topically active corticosteroids include, but are not limited to, bellcromethasone 17,21-dipropionate, alclometasone dipropionate, budesonide, 22S budesonide, 22R budesonide, But are not limited to, monopropionate, clobetasol propionate, diflorosone diacetate, fluney solids, plurandrelinolide, fluticasone propionate, halobetasol propionate, ≪ / RTI > and triamcinolone acetonide. In a preferred embodiment of the present invention, the locally active corticosteroid is bellcloomethsone diproprionate. The effective amount of locally active corticosteroids in each dosage form can vary from patient to patient and can be readily determined by those skilled in the art by well known dose-response studies. Such effective amounts will generally range from about 0.1 mg / day to about 8 mg / day, and more typically from about 2 mg / day to about 4 mg / day.

It is yet another object of the present invention to provide a method of alleviating gastrointestinal, hematopoietic and subcutaneous effects of acute radiation syndrome in a subject in which a high transmittance dose of radiation is contained in a substantial part of the body of the subject in a short period of time.

It is yet another object of the present invention to provide a method for treating acute radiation syndrome in a subject in which a high transmittance dose of radiation is received in a substantial portion of the body of the subject in a short period of time, Or its metabolite, and the oral dosage form is effective for topical or local treatment of the gastrointestinal tract and oral cavity of the subject, and further the subject is a subject of the symptoms of inflammation due to tissue damage resulting from radiation or chemotherapy treatment And to provide a method of representing a target chain. Representative topically active corticosteroids include, but are not limited to, bellcromethasone 17,21-dipropionate, alclometasone dipropionate, budesonide, 22S budesonide, 22R budesonide, But are not limited to, monopropionate, clobetasol propionate, diflorosone diacetate, fluney solids, plurandrelinolide, fluticasone propionate, halobetasol propionate, ≪ / RTI > and triamcinolone acetonide. In a preferred embodiment of the present invention, the locally active corticosteroid is bellcloomethsone diproprionate. The effective amount of locally active corticosteroids in each dosage form can vary from patient to patient and can be readily determined by those skilled in the art by well known dose-response studies. Such effective amounts will generally range from about 0.1 mg / day to about 8 mg / day, and more typically from about 2 mg / day to about 4 mg / day.

Yet another object of the present invention is to provide a method of treating an infection (e. G., A microbial infection) in a subject by administering to the subject a peptide comprising or comprising the amino acid sequence of Table 1 or an analog, derivative or variant thereof or a clear chemical equivalent thereof And / or preventing the disease. As an example, the subject may be infected or at risk of being caught. In one embodiment, the peptide modulates congenital immunity in a subject to treat and / or prevent infection in the subject.

Exemplary infections that can be treated and / or prevented by the methods of the present invention include infections by bacteria (e.g., gram-positive or gram-negative bacteria), infections of fungi, infections by parasites, and infections by viruses . In one embodiment of the invention, the infection is a bacterial infection (e . G. , E. coli , Klebsiella pneumoniae , Pseudomonas aeruginosa, Infections caused by aeruginosa , Salmonella spp ., Staphylococcus aureus , Streptococcus spp ., and vancomycin-resistant Enterococcus ) to be. In yet another embodiment, the infection is a fungal infection (e. G., Infection by a mold, yeast, or higher fungus). In yet another embodiment, the infection is a parasitic infection (e.g., jiahreu dia duo Gardena lease (Giardia infection by single-cell or multicellular parasites, including, for example, duodenalis , Cryptosporidium parvum , Cyclospora cayetanensis , and Toxoplasma gondii . In still yet another embodiment, the infection is selected from the group consisting of viral infections (e. G., AIDS, avian flu, chickenpox, herpes simplex, common cold, gastroenteritis, glandular fever, influenza, measles, mumps, , SARS (SARS), and viruses associated with lower or upper respiratory tract infections (e. G., Respiratory syncytial viruses). Dosage formulations.

The dosage form of RIVPA (SEQ ID NO. 5) is an aqueous aseptically treated sterile solution for injection. Each vial contains 5 mL of a 60 mg / mL solution (300 mg of RIVPA (SEQ ID NO. 5)). RIVPA (SEQ ID NO. 5) is formulated in infusion water and the pH is adjusted to a target value of 6.0. The formulation does not contain excipients and the osmolality is ~ 300 mOsm / kg.

Route of administration

RIVPA (SEQ ID NO. 5) The drug product will be diluted in sterile saline at an appropriate concentration for injection and is measured in mg / kg based on the weight of the recipient and the designed dose level. Diluted RIVPA (SEQ ID NO. 5) will be administered as intravenous (IV) infusion at 25 mL over 4 minutes once every 3 days.

Example

Peptide synthesis

The peptides of Table 1 were synthesized using solid phase peptide synthesis techniques.

All of the required Fmoc-protected amino acids were weighed in triple molar excess relative to 1 millimole (mmol) of the desired peptide. The amino acids were then dissolved in dimethylformamide (DMF) (7.5 ml) to prepare 3 mMol of solution. Appropriate amounts of Rink amide MBHA resin were weighed in consideration of resin substitution. The resin was then transferred to an automated synthesizer reaction vessel and pre-immersed for 15 minutes with dichloromethane (DCM).

To the resin was added 25% piperidine in DMF (30 ml) and mixed for 20 minutes to deprotect the resin. After deprotection of the resin, a solution of 3 mMol of amino acid was added to a solution of 4 mMol of 2- (1H-benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) The first coupling was carried out by mixing with N, N-diisopropylethylamine (DIEPA). The solution was pre-activated for 5 minutes before adding to the resin. The amino acid was allowed to couple for 45 minutes.

After coupling, the resin was thoroughly rinsed with DMF and dimethylacetamide (DMA). The attached Fmoc-protected amino acids were deprotected in the same manner as described above and the following amino acids were attached using the same coupling scheme AA: HBTU: DIEPA.

After completion of the synthesis, the cleavage cocktail containing 97.5% trifluoroacetic acid (TFA) and 2.5% water was used to strip the peptide from the resin. The resin was soaked for 1½ hours in the cutting cocktail. The solution was then filtered by gravity using a Buchner funnel and the filtrate was collected in a 50 ml centrifuge tube. The peptides were separated by precipitation with cold diethyl ether. After centrifugation and diethyl ether decanting, the crude peptide was washed once more with diethyl ether before drying in a vacuum desiccator for 2 hours. The peptides were then dissolved in deionized water (10 mL), frozen at -80 DEG C and lyophilized. Dried peptides for HPLC purification were then prepared.

Due to the hydrophilic nature of these peptides, diethyl ether peptide separation did not perform well. Therefore, chloroform extraction was required. TFA was evaporated and the resulting peptide residue was dissolved in 10% acetic acid (15 mL). The acetic acid peptide solution was washed twice with chloroform (30 ml) to remove impurities and scavengers from the solution. The peptide aqueous solution was then frozen at -80 DEG C and lyophilized to generate the pulverized peptide prepared for HPLC purification.

Each of the peptides + RIxVPA (SEQ ID NO. 33) and + RIVPAx (SEQ ID NO. 34) contained one N-methyl amino acid. This coupling was carried out by mixing together N-methyl amino acid, PyBroP and N-hydroxybenzotriazole * H2O (HOBt) and DIEPA solution in a resin containing RV. After being allowed to couple for 45 minutes, the N-methyl amino acid was then re-coupled to ensure complete coupling. It was observed that the coupling after the N-methyl amino acid was not completely complete. This coupling was therefore carried out using N, N, N ', N'-tetramethyl-O- (7-azabenzotriazol-1-yl) uronium hexafluorophosphate (HATU) instead of HBTU . Which still produced crude peptides typically containing two impurities compiled to 30-40% of the total purity. The peptides were purified under modified HPLC conditions to separate pure peptide peaks from closely eluting impurities.

R (tBg) V1KR (tBg) V2 (SEQ ID NO. 91) is an 8-residue peptide dendrimer with a symmetrical branching occurring in a fourth amino acid lysine with two amine functional groups. Peptides were synthesized by solid-phase peptide synthesis techniques using a di-Fmoc protected fourth amino acid to facilitate coupling of branches using the general synthetic techniques described above.

In addition, these peptides can also be synthesized using solution phase peptide synthesis techniques (Tsuda et al. 2010) and are commonly known to those skilled in the art.

oral- In mucositis  efficacy

RIVPA (SEQ ID NO. 5) and other IDRs modulate the innate defense response to tissue injury, reducing the severity of damage caused by the continuous inflammatory process and improving disease resolution. This contribution of IDR has been demonstrated in chemotherapy-induced oral and GI mucositis in mice, radiation-induced oral mucositis in hamsters and DSS-induced colitis in mice. In each of these models, initial injury is believed to trigger a sequential step of congenital defensive signaling that increases the severity of injury (Marks 2011; Sonis 2010). RIVPA (SEQ ID NO. 5) and other IDRs counteract the signaling sequential steps, reducing the severity of injury induced and reducing the duration of severe tissue injury.

Optimal dosing regimens for RIVPA (SEQ ID NO. 5) and other IDRs identified in the MRSA bactericidal model were further confirmed in the injury model, where a longer period of disease is more beneficial for repeated dosing. Doses of 25 mg / kg every 3 days were found to be optimal, indicating that despite rapid PK removal (within minutes) of RIVPA and other IDRs (SEQ ID NO. 5) from the circulation of the mice, their continued pharmacodynamic Reflect the impact.

RIVPA (SEQ ID NO. 5) significantly reduced the severity and duration of mucositis in a radiation-induced oral mucositis model of hamsters, especially when administered every three days during fractional irradiation therapy. These studies confirm that optimal dosing of RIVPA (SEQ ID NO. 5) involves administration every 3 days and that a dose level of 25 mg / kg is effective. In this model, canine male Golden Syrian hamsters were treated with 7.5 Gy of radiation toward the inverted left ball pocket at 0, 1, 2, 3, 6, 7, 8 and 9 days. Mucositis was assessed every 2 days between 7 and 35 days, with peak mucositis severity occurring generally on about 19 days. In the first study, RIVPA (SEQ ID NO. 5) (25 mg / kg IV) was administered every 3 days starting on day 0 and continuing until day 33 (Q3d d0-33) 1, 2, 3, 4, 7, 8, 9 days) or 6 days to 24 days (Q3d d6-24). On the day that both RIVPA (SEQ ID NO. 5) and radiation were administered, RIVPA (SEQ ID NO. 5) was given 2 hours post radiation. The results of these studies are shown in Fig. Treatment with RIVPA (SEQ ID NO. 5) was most effective when administered every 3 days or on the day of radiation over the radiation period, whereas treatment starting on the 6th day after radiation initiation was not beneficial (i.e., Q3d d6-24) . Follow-up studies were performed evaluating the administration of Q3d d0-33 at 25 mg / kg IV RIVPA (SEQ ID NO: 5) every 3 days (ie, 0, 3, 6 and 9 days) . The results of these studies are shown in FIG. Treatment every 3 days during radiation was found to be optimal and reflects the persistence of the RIVPA (SEQ ID NO. 5) pharmacodynamic effects associated with the reduction of injection stress caused by less IV infusion in these small rodents Seems to be.

RIVPA (SEQ ID NO. 5) also showed efficacy in mouse models of chemotherapy-induced oral and gastric mucositis, consistent with the response of innate immune responses to chemotherapy and / or radiation injury. In these studies, administration of RIVPA (SEQ ID NO. 5) was associated with a statistically significant reduction in the duration of severe oral mucositis in the chemotherapy-induced mucositis model in mice. Although mild GI injury in the control group provided statistically insignificant results, a trend toward reduced colitis was also observed. In each study, 5-fluorouracil (60 mg / kg IP) was administered to male C3H / HeN mice at -4 and -2 days. At day 0, a chemical laceration was applied beneath the mouse tongue to induce mucositis, which generally peaked at 2 days. The mouse tongue was graded for mucositis daily for 1 to 14 days, grade ≥3 indicating severe mucositis. Body weight was also measured daily and colitis severity was measured by video endoscopy at 4 and 7 days. In the first study, RIVPA (SEQ ID NO. 5) (25 mg / kg IV) was administered once-four days immediately prior to chemotherapy, two times on days -4 and -2 immediately after chemotherapy, Lt; / RTI > RIVPA (SEQ ID NO. 5) administration was most effective (i. E., -1, 2 and 5 days) at various timepoints during the peak mucosal injury period. The results of this study are shown in Fig. In a second study, RIVPA (SEQ ID NO. 5) (25 mg / kg IV) was administered at -1, 2 and 5 days, -1, 1 and 3 days or 0, 2 and 4 days. The results of this study are shown in FIG. Statistically significant changes in the severity of severe mucositis (FIG. 5 - Panel A), severity of colitis at 4 days (FIG. 5 - Panel B) and mean weight loss (FIG. 5 - Panel C) were correlated among the groups. In a third study, RIVPA (SEQ ID NO. 5) (25 or 5 mg / kg IV as indicated) was administered at days -1, 2 and 5 or at days 1 and 3. Also, dosing regimens with RIVPA (SEQ ID NO. 5) every 3 days were most effective, and reduced dose levels resulted in reduced efficacy. Mucositis, colitis and weight results from this study are shown as A, B and C, respectively, in FIG. Statistical significance was assessed for oral mucositis using a chi-square test and for the curve under-weight area (AUC) using ANOVA for rank. R (tBg) V1KR (tBg) V2 (SEQ ID NO. 91) and 92 also demonstrated efficacy in a mouse model of chemotherapy-induced mucositis where the mucositis grade was assessed for 4 days after induction of mucositis 16, Fig. 17). Administration of a therapeutic agent with R (tBg) V1KR (tBg) V2 (SEQ ID NO. 91) and RIV (mp2) A-NH2 (SEQ ID NO. 92) at -1 and 2 days with a dose of 25 mg / kg IV Respectively.

radiation Efficacy in response to injury

RIVPA (SEQ ID NO. 5) and other IDRs modulate the innate defense response to tissue injury, reducing the severity of damage caused by the continuous inflammatory process and improving disease resolution. As described above, IDR can alleviate the response to radiation damage in the oral mucositis model (Figs. 1 and 2). In another model, in a prophylactic evaluation of radiation-induced mucositis (25 Gy administered to mouse nose at day 0), RIVPA (SEQ ID NO. 5) (5 doses of 25 mg / kg IV administered every 2 days Administration) had no significant effect on disease progression. Progressive thinning of the mouse tongue was observed at 0, 2, 4, 6, 8, and 10 by histopathological analysis of basal and early hypoproteotics, mitosis, and total epithelial cell count per unit area and unit length Day. It is noted that the dose of radiation used (25 Gy) was chosen so that progressive thinning of the tongue epithelium was observed but no explicit mucositis occurred. These results demonstrate a deficiency in the proliferative potential of RIVPA (SEQ ID NO. 5), and the effect of RIVPA (SEQ ID NO. 5) is only observable if the pathway is stimulated by explicit tissue damage or pathogenesis Prove that.

Efficacy in gastrointestinal tract

The ability to directly protect the GI mucosal surface of prophylactically or therapeutically administered IV RIVPA (SEQ ID NO. 5) was identified in a DSS-induced colitis model. In this model, DSS was administered as a 3% DSS solution in drinking water of male C57BL / 6 mice on days 0-5 of study. Colitis was monitored by video endoscopy on days 7, 14, and 21. (Q3d d0-18), 3-18 days (Q3d d3-18), or 6-18 days (Q3d d6-18), respectively, in the presence of RIVPA (SEQ ID NO. Lt; / RTI &gt; every 3 days. The results of the study are shown in Fig. By day 14, all RIVPA (SEQ ID NO. 5) treatment regimens demonstrated a statistically significant reduction in endoscopic colitis severity grade. However, the reduction in grade at 7 days is due to the group receiving at least two doses of RIVPA (SEQ ID NO. 5) up to that time (i.e., Q3d d0-18 and Q3d d3-18, but not Q3d d6-18 ). At day 21, all three treatment groups appeared to respond in a similar manner. On day 21, the histopathology of the colon showed statistically significant reductions in edema and necrosis in some RIVPA (SEQ ID NO. 5) -treated groups, while other RIVPA (SEQ ID NO. 5) Which is similar to a reaction that does not reach. Statistical analysis was performed using t-test, with asterisk showing a statistically significant difference from the control (p <.05).

As described above, IDR can also reduce the duration and / or severity of gastrointestinal mucositis in chemotherapy-induced mucositis models (Figs. 4, 5, 6).

Efficacy in Infected Animals

RIVPA (SEQ ID NO. 5) reduces bacterial burden and improves survival in the presence or absence of antibiotic treatment in a variety of murine infection models and provides consistent efficacy at dose levels of 25 mg / kg and greater And a sustained pharmacodynamic effect of less than 5 days. RIVPA (SEQ ID NO. 5) efficacy complements antibiotic therapy in both normal and immunocompromised mice. The efficacy of RIVPA (SEQ ID NO. 5) has been demonstrated for diseases caused by gram-positive (S. aureus and MRSA) and gram-negative (Klebsiella, E. coli and B. pseudomalai) .

S. Aureus

RIVPA (SEQ ID NO. 5) was evaluated both as standard-alone treatment and in combination with vancomycin treatment.

RIVPA (SEQ ID NO. 5) treatment increased survival in the MRSA peritoneal infection model when administered in combination with the antibiotic dose of vancomycin (Study #: D-7-E-11). RIVPA (. SEQ ID NO 5) (50 mg / kg) or saline treatment MRSA (UC6685; 8.2 x 10 ⁷ colony forming units [cfu]) female to vaccination before 48 or 72h to CF-1 mice (N = 10 / Group). Vancomycin treatment (3 mg / kg) was administered subcutaneously (SC) at 1 and 5 h post-infection. Survival was monitored once a day for 5 days. The results of these studies are shown in Fig.

RIVPA (SEQ ID NO. 5) is also effective when administered by itself. Multiple studies with IV-administered RIVPA (SEQ ID NO. 5) were performed in MRSA bacteremia models. Administration of RIVPA (SEQ ID NO. 5) demonstrated dose response in this model of T-cell deficient immunocompetent Balb / c mice or nu / nu mice, with a single dose of 50 mg / kg compared to saline control Induced increased survival. In the first study, MRSA (USA 300, 7.3 log10 cfu) was administered via IV infusion into the tail vein of female Balb / c mice at 0 hr. At 4 hours prior to infection, a single dose of saline or RIVPA (SEQ ID NO. 5) was injected intravenously into the tail vein at the indicated dose level. A second-line antibiotic treatment (linezolid, 6.25 mg / kg) was given orally once daily immediately after infection. Survival was monitored for 21 days after infection. The results of this study are shown in FIG. In a second study, RIVPA (SEQ ID NO. 5) (IV) or saline (IV) was administered once through the tail vein into female nu / nu mice at 4 h prior to infection with MRSA (strain USA300, 7.0 log ₁₀ cfu) Respectively. Survival was monitored for 14 days as shown in Fig. A statistically significant difference in survival (ie, p ≤ 0.05) was found at a dose level of 50 mg / kg when assessed using the Kaplan-Meier analysis of each treatment group compared to the saline control.

In summary, a study using a single IV RIVPA (SEQ ID NO. 5) treatment in various S. aureus infection studies demonstrated the following:

The effect of RIVPA (SEQ ID NO. 5) is dose dependent at 1-50 mg / kg mice and dose levels of 25 mg / kg and higher consistently demonstrate efficacy (Table 2; Figures 9 and 10 ). These dose levels were also effective in the more chronic disease conditions available in the injury model (Figures 1-6).

Table 2: Successful treatment rate of S. aureus infection with single IV RIVPA (SEQ ID NO. 5)

 (i) Successful treatments have demonstrated at least a 20% increase in survival compared to the relevant saline control.

(ii) Study #: TPS-8-B-100, TPS-8-B-150, TPS-8-

(iii) Study #: TPS-8-B-100, TPS-8-B-150, TPS-8-

B-114, TPS-8-B-100, TPS-8-B-100, TPS-8- B-101

(v) Study #: TPS-8-B-100, TPS-8-B-150

Daily administration of RIVPA (SEQ ID NO. 5) is not necessary and administration every 2 days or every 3 days is sufficient. In a more chronic disease situation available in the Shanghai model, it was additionally confirmed that every 3 days administration appears to be optimal (data not shown).

RIVPA (SEQ ID NO. 5) can be administered up to 24 h after the onset of infection in the MRSA bactericidal model and still provide survival benefit (data not shown). Thus, its action is rapid.

Depending on the dose level, a single dose of RIVPA (SEQ ID NO. 5) can be administered up to 5 days prior to the start of infection, still providing a survival advantage (data not shown) Reflects the sustained pharmacodynamic effects of RIVPA (SEQ ID NO. 5) despite pharmacokinetic (PK) elimination (within minutes).

The survival benefit provided by the RIVPA (SEQ ID NO. 5) therapeutic agent can last for at least 21 days (FIG. 9).

RIVPAY ^* (SEQ ID NO. 90) and R (tBg) V1KR (tBg) V2 (SEQ ID NO. 91) (administered at 5 mg / kg before 4 hours of infection) also improve survival in MRSA bacteraemia models (Figs. 18 and 19).

Topical administration of RIVPA (SEQ ID NO. 5) has also proven effective when the administration is localized to the site of infection. In the Gram-positive peritoneal infection model of mice, RIVPA (SEQ ID NO. 5) significantly reduced bacterial load by over 7 logs (Study #: D-7-E-14). Peritoneal (IP) injections of S. aureus (Cat. No. 25923, ATCC, 6 × 10 ⁷ cfu) into 5% mucin were IP-administered with female CD-1 mice (N = 8 / RIVPA (SEQ ID NO. 5) (9.5 mg / kg) was injected IP after 4 h. Mice were sacrificed at 24h post infection and peritoneal lavage fluid was evaluated for bacterial count. The results of this study are shown in Figure 11 (each data point represents the result from an individual mouse - a dead mouse is given the highest bacterial count of any mouse acquired in the study and is represented by an open symbol in the graph) .

RIVPA (SEQ ID NO. 5) also significantly reduced bacterial load in neutropenic mice in the S. aureus thigh abscess infection model when administered as topical intramuscular (IM) infusion. Female Swiss albino mice (N = 8 / group) were treated with Cp (100 mg / kg) on days 3 and 1 before IM infection with S. aureus (catalog number 29213, ATCC, ~ 9.5 x 10 ⁵ cfu) Neutrophils were decreased. RIVPA (SEQ ID NO. 5) (50 mg / kg) was administered IM for 24 h before infection and SC was administered at 1, 6 and 18 h after infection with vancomycin (100 mg / kg). The number of bacteria cfu present in infected thighs was assessed at 24h after initiation of infection in each group. The results of this study are shown in Fig.

Klebsiela

RIVPA (SEQ ID NO. 5) increased survival in Gram-negative peritoneal infection models when administered topically (IP) or systemic (IV). For reference, systemic administration appears to be better or better than topical administration. RIVPA (SEQ ID NO. 5) Therapeutic agent (24 mg / kg) was injected intravenously in a dose of 2.8 x 10 ⁵ cfu (Figure 13 - Panel A) or 5.3 x 10 ² cfu (24 h before infection or 4 h after infection) or IV (4 h after infection) were administered to female Balb / c mice (N = 8 / group) The protective effect of RIVPA (SEQ ID NO. 5) in this situation is shown in FIG. Survival endpoints for more bacterial inoculum-administered animals are shown (panel A). All animals dosed with fewer inoculations survived in all groups (Panel B) and evaluated at 24 h post-infection for clinical signals (eg, nape, reduced motion, curved abdomen, etc.); These were summarized as clinical grades.

Efficacy in skin damage

Systemically administered RIVPA (SEQ ID NO. 5) was also effective in skin injuries and infections and accelerated skin healing in the MRSA skin infection model. Infection was initiated on day 1 after removing hair from the back area of each mouse. RIVPA (SEQ ID NO. 5) (25 mg / kg IV or 100 mg / kg SC) was administered at 4 h before infection and at various time points after infection as indicated. Oral linezolid was used as a comparative factor and administered daily at 12.5 mg / kg. Each mouse was anesthetized and shaved using isoflurane at day 0 (-1h), and the skin was injured by seven consecutive application and removal of surgical tape. This lesion was then immediately infected by topical administration of 10 μL of bacterial suspension and delivered 7.6 log ₁₀ cfu of total irritant per mouse. By measuring the bacterial burden in the punch biopsy of the skin at 48 h (FIG. 14-Panel A) and 96 h (FIG. 14-Panel B) after bacterial stimulation and after 48 h (Fig. 14-Panel D), the efficacy was assessed by macroscopic evaluation of the digital image of the skin by a blinded pathologist. For reference, localizations of any isolated bacteria (i.e., on or on the skin surface) were not measured, but neither linazolid or RIVPA (SEQ ID NO. 5) was found in biopsies at 48 or 96 h But did not reduce bacterial load. Nonetheless, wound healing was evident. The mean bacterial burden in each treatment group was statistically compared to that of its time-matched, saline control group, using a t-evaluation comparison of the averages that performed in Excel and estimated aequality variability. Comparisons returned with a p-value ≤ 0.05 were considered statistically different.

Safe pharmacology in healthy animals:

Two pilot and two final repeat-dose toxicity studies were performed with RIVPA (SEQ ID NO. 5) in mice and cynomolgus monkeys using intravenous (IV; slow bolus) administration route . All studies were performed by LAB Research Inc. (Canada).

Non-GLP pilot toxicology studies showed that the maximum tolerated dose (MTD) of a single dose of RIVPA (SEQ ID NO. 5) administered with IV infusion over 30 to 60 seconds was 88 mg / kg (actual dose) in mice . In a non-GLP pilot study of nonhuman primate (NHP), a dose of 90 (1 animal), 180 (both animals) and 220 (1 animal) mg / kg RIVPA (SEQ ID NO. 5) Light clinical signals (shallow breathing / dyspnea, reduced activity, partially coiled eyes and muscle spasms) were recorded in one or both animals after administration. They disappeared within minutes without detectable residual effect.

The safety of multiple daily injections of RIVPA (SEQ ID NO. 5) was also evaluated in GLP studies in mice and cynomolgus monkeys. In mice, doses of 20, 60, or 90 mg / kg / day were given for 14 days IV. Lethal doses were observed at high doses, predominantly with dyspnea and lateral infections. Other animals were also observed in this dose at a dose of 60 mg / kg, but no clinical signals were shown at this dose. At 20 mg / kg no evaluation documentation-related mortality or clinical signs were observed. In all groups of live animals, there was no evidence of toxicity in any organ or abnormal biochemistry or hematology. Side effects were not observed at 20 mg / kg for 14 days.

RIVPA (SEQ ID NO. 5) at 20, 80 and 160 mg / kg / day was given IV to cynomolgus monkeys for 14 days. Transient reduced activity and partially coiled eyes were observed continuously in most animals during and immediately after administration at 160 mg / kg for the first 3 days and then sporadically over the rest of the dosing period. In all cases, these clinical signals disappeared within minutes. Adverse effects were not observed for any other measured variables or by microscopy in any tissue. Administration of RIVPA (SEQ ID NO. 5) at doses of 20 and 80 mg / kg / day resulted in no evidence of any toxicity. A dose level of 80 mg / kg / day was considered a No-Observed-Effect-Level (NOAEL) in this study.

The effect of RIVPA (SEQ ID NO. 5) was not observed in the central nervous system (CNS) in any study at any dose level and was shown to be dose-dependent for radiation-labeled RIVPA (SEQ ID NO. 5 ) Were rarely observed in mouse CNS. No interaction between the CNS receptor and ion channel battery and RIVPA (SEQ ID NO. 5) was detected in vitro.

Cardiovascular (CV) / lung studies in cynomolgus monkeys did not show cardiovascular effects or changes in ECG parameters using a single IV dose of 20 or 80 mg / kg. No respiratory effects were observed at doses of 20 or 80 mg / kg. At a dose of 80 mg / kg of this study, RIVPA (SEQ ID NO. 5) was associated with transient sagging eyelids and draining during dosing. Administration of RIVPA (SEQ ID NO. 5) at 220 mg / kg was associated with transient severe clinical signs such as drooping eyelids, tremor, burning, pale, spasms and stenosis. In 1 animals, high dose resulted in a pronounced decrease in respiratory rate, resulting in bradycardia, hypotension, and lethality.

In general, NOAELs are considered to be 80 mg / kg / day for cynomolgus monkeys, since transient clinical signals are limited to a single study and only occurred in only two of the 98 doses of drug at this dose level.

A carcinogenic, mutagenic or reproductive toxicity study was not performed with RIVPA (SEQ ID NO. 5).

The effect of RIVPA (SEQ ID NO. 5) on the innate defense system is highly selective. Consistent with this finding, no changes were observed in immuno-associated organ weights, histopathology, hematology, and clinical chemistry during the mouse and NHP 14 -yl toxicity studies. In subsequent studies, no effect on T-cell, B-cell or NK-cell number was observed after 14 days of administration of intravenous RIVPA (SEQ ID NO. 5) in NHP. RIVPA (SEQ ID NO. 5) did not promote the proliferation of mouse or human normal blood cells or the proliferation of primary human leukemia cells in vitro in vitro. Overall, it does not show the potential for RIVPA (SEQ ID NO. 5) resulting in immunotoxicity or non-specific immune activation. No other effects on the phenotypic expression of cells associated with excessive activation or inhibition of the adaptive immune response or adaptive immunity after administration of RIVPA (SEQ ID NO. 5) were detected.

In summary, the major toxicological findings are acute-onset respiratory depression with transient dyspnea, lateralization and transient depressed activity. In most of its severity, acute toxicity caused lethality. Clinical signals are all reversible when dosing is discontinuous, and animals have been observed to recover within minutes without clinical signs or subsequent detrimental sequelae of toxicity results. Cardiovascular / pulmonary safety pharmacological studies in nonhuman primates confirmed that no cardiac toxicity or QT syndrome occurred.

Observed respiratory depression occurred at various dose levels in various species and was not predicted by relative growth scaling. In particular, mice rarely develop at 60 mg / kg (HED: ~ 5 mg / kg) and appear to be the most sensitive species with acute toxicity generally occurring at 90 mg / kg (HED: ~ 7 mg / kg). Acute toxicity occurred occasionally at 160 mg / kg (HED: ~ 50 mg / kg) and persisted at 240 mg / kg (HED: ~78 mg / kg), as opposed to NHP (cynomolgus monkey) Further studies with RIVPA (SEQ ID NO. 5) analogs in acute mouse toxicity studies have shown that toxicity is associated with charge rather than with the specific structure (amino acid sequence) or the target protein binding state of the molecule, And that it is due to the high instantaneous concentration of charged molecules scaling to the blood volume as received. In addition, mechanistic studies in mice have shown that dyspnea is due to altered activity of the transverse nerve.

Safety against leukopenia and / or infection Pharmacology:

In a non-GLP pharmacology study, RIVPA (SEQ ID NO. 5) did not alter the recovery of circulating blood cell populations after induction of leukopenia in CD-1 mice. Leukopenia was induced by 2 IP infusion of Cp (150 mg / kg at 1 day and 100 mg / kg at 4 days), leading to well-established leukopenia until 4 days, which lasted for approximately 10 days. Saline or RIVPA (SEQ ID NO. 5) (20 or 50 mg / kg) was administered IV on days 5, 7, 9 and 11. Six animals per group were sacrificed at 6, 8, 10, 12 and 14 days, respectively, and evaluated for complete blood counts and differences. None of the levels or dynamics of total leukocyte and leukocyte percentages were altered during the recovery process when compared to the vehicle control (Figure 15).

Infection studies in leukopenia-reduced animals have shown that RIVPA (SEQ ID NO. 5) does not interfere with antibiotic efficacy.

(SEQ ID NO. 5) which is treated by CYP450 enzyme or by inhibition of CYP450 enzyme, primary metabolism of RIVPA (SEQ ID NO. 5) by protease across body tissue and RIVPA ) Removal suggests that pharmacokinetic drug-drug interactions will be minimized. &Lt; Desc / Clms Page number 2 &gt;

iv. Clinical experiment

Clinical trials with RIVPA (SEQ ID NO. 5) were obtained in a Phase 1 study. The primary objective of the study was to determine the maximum tolerated dose (MTD) of single and repeated dose increments of injectable solutions of RIVPA (SEQ ID NO. 5) after IV administration in healthy volunteers. The secondary objective of this study was to evaluate the capacity-limiting toxicity (DLT), safety, PK and pharmacodynamic (PD) profiles of RIVPA (SEQ ID NO. 5) after single and repeated increases IV dose of RIVPA (SEQ ID NO. 5) . This study was divided into two phases: Single-Incremental Capacity (SAD) and Multi-Incremental Capacity (MAD).

Human safety

Single IV dose of RIVPA (SEQ ID NO. 5) was well tolerated up to the maximum evaluated dose (8 mg / kg), and daily IV dose was well tolerated up to the maximum evaluated dose (6.5 mg / kg for 7 days). There was no dose-limiting toxicity (DLT), and MTD was not reached at any one time in the test. There were no mortalities reported during the study and there were no clinically significant, severe, or serious adverse events (Adverse Events) (AEs). No significant differences in safety concerns or mean values or changes from baseline in vital sign measurements, clinical laboratory or electrocardiogram (ECG) results between the drug treated and placebo control subjects were observed.

Single Increased Capacity Face:

The occurrence of TEAE in such a subject to which RIVPA (SEQ ID NO. 5) has been administered is not dose-related and is clinically significant in subjects receiving RIVPA (SEQ ID NO. 5) as compared to the subject to which the placebo is administered The incident did not occur at a significantly higher rate. The most frequently reported TEAE (observed in more than one subject administered RIVPA (SEQ ID NO. 5) and at a higher percentage (%) than the placebo subject) For example, vascular puncture hematoma, vascular puncture site reaction, and vascular puncture site pain. All vascular hole-related events were weak and were not associated with the QI-mediated study treatment. The second most frequently reported TEAE is the Nervous System Disorder, especially headache and dizziness; These events are only mild to moderate. All other TEAEs were reported by only one subject at any given dose level (maximum of 3 dose levels). No clinically significant trends in the natural or metamorphic nature of TEAE have been demonstrated in any study population.

Multiple Increased Capacity Pace:

The highest incidence of TEAE was observed at the two highest dose levels (4.5 and 6.5 mg / kg / day). The occurrence of "possibly-related" events was also higher at the two highest dose levels. However, due to the smaller sample size (4 subjects treated with the active agent in each group), it was impossible to conclude whether the results clearly showed dose-response. Most TEAEs were not associated with study treatment, were weak in severity, and only one case was reported to be moderate. The most frequently reported TEAEs in subjects to which RIVPA (SEQ ID NO. 5) has been administered include General Disorders and Administration Site Conditions (i.e., procedure-related events) such as vascular hematoma, Vascular puncture site reaction and vascular puncture site pain. All vascular puncture-related events were weak and were not associated with treatment. Increased alanine aminotransferase (ALT) and back pain were reported by 3 (15.0%) subjects receiving RIVPA (SEQ ID NO. 5), and these events were reported in only one (10.0% Lt; / RTI &gt;

human Pharmacokinetics

After IV administration in human subjects consistent with findings in animal studies, RIVPA (SEQ ID NO. 5) is removed from circulation within minutes. Rapid elimination of RIVPA (SEQ ID NO. 5) at the single-dose face of a healthy volunteer Phase 1 trial reduced plasma levels to less than 10 percent of the maximum concentration (Cmax) within 9 minutes after the initiation of 4-min IV infusion . After a rapid decline, a slower removal phase was observed. The average time of maximum concentration (Tmax) after initiation of injection at the dose range of 0.15 mg / kg to 8 mg / kg ranged from ~ 4 minutes to 4.8 minutes. The maximum plasma concentration and total exposure levels were dose-proportional and the removal of RIVPA (SEQ ID NO. 5) from the circulation was consistent with the mouse and NHP experiments.

Taking into consideration the high elimination and the short elimination half-life, accumulation after daily injection was not expected to occur. In a multi-dose Phase 1 study, RIVPA (SEQ ID NO. 5) was administered daily for 7 days and the pre-dose concentration measured at 8 days (24h after beginning injection at 7 days) as well as days 5, 6, 7 Below the lower limit of quantitation (LLOQ) for all subjects.

human Pharmacodynamics

In an in vitro study using a blood sample obtained during the Phase 1 healthy human volunteer study, a number of cytokines and chemokine assays were quantified with LPS 4 hours after in vitro stimulation of whole blood. Interpersonal variability at the analyte level is greater than the response to RIVPA (SEQ ID NO. 5) or placebo administration or any variation in time, thus the data are self-normalized using individual pre-dose analyte levels All responses to each individual subject were normalized (activity ratio). The effect of RIVPA (SEQ ID NO. 5) on analyte activity ratio (AR) is not constant or linearly dose-responsive over time. Nevertheless, at doses ranging from 0.15 to 2 mg / kg, an increase in "anti-inflammatory status" was evidenced (i.e., higher levels of TNFα and IL- Anti-inflammatory TNF RII and IL-1ra levels).

b. IDR  Scientific Basis for Injection

Mucositis

Mucositis is associated with a concomitant dysregulation of the congenital defense system, which results in a sequential step of inflammatory action which further compromises the mucosal lining and leads to obvious mucositis (Sonis, 2004). In particular, chemotherapy or radiotherapy causes damage to the underlying endothelium and epithelium, whereas the response of the congenital defense system to the resulting "DAMPS" results in a continuous inflammatory process, which worsens the damage. Recent studies evaluating gene expression in animals and humans with severe oral mucositis propensity supported the role of congenital defense systems in the disease (Sonis, 2010). In addition, sub-gastrointestinal mucositis also results from a similar mechanism (Bowen, 2008).

Acute radiation syndrome

Acute radiation exposure is associated with damage to epithelium (skin), bone marrow (hematopoietic syndrome), and gastrointestinal tract (GI). Furthermore, as radiation exposure increases, the mortality rate becomes increasingly acute and limits the potential for therapeutic intervention. Initial mortality after acute radiation exposure (<2 weeks) is associated with damage to the gastrointestinal tract. Acute radiation causes direct damage to stem cells in the base of the river queen, which results in mitosis through mitotic arrest and apoptotic mechanisms (Potten 1997a, Potten 1997b). Recovery of these impairments and / or long-term sequelae have been shown to be associated with both GI microbial and congenital immune recovery responses (Crawford 2005; Garg, 2010). Progressive studies of normal tissue and tumorigenic radiation biology have demonstrated a significant role for the response of the innate immune system to radiation (Schaue and McBride, 2010; Lauber 2012; Burnette 2012). In addition, agonists targeting the innate defense system (i.e., TLR-9 agonists) have been shown to be radiation protective in the GI ARS setting (Saha, 2012).

As a result of radiation tumor therapy, oral and GI mucositis act as proxies in the GI component of ARS. Consistent with the role of congenital defenses in mucositis and the function of IDR, efficacy with IDR has been demonstrated in many mucosal injury models. In particular, studies in both chemotherapy and radiation-induced oral and gastro-intestinal mucositis have shown that IDR can reduce the peak intensity and duration of mucositis, leading to ~ 50% reduction in the duration of severe mucositis 1, 2, 4, 5, 6, 7, 8, 16, 17).

In addition, consistent with the IDR effect on the host's innate immunity, efficacy in the infectious model was significantly reduced in both immune (e. G., FIG. 9) and immunocompromised (leukocyte depletion or T- And gram-positive (Figures 9, 10, 18, 19) bacterial pathogens.

The IDR is administered systemically and affects the skin as well as the mucosal surface (e.g., oral mucosa, colon [Fig. 3]) and can be affected by both the whole body (Figs. 7, 8, 9 and 10) effective.

In summary, IDR mediates the innate defense response to injury (Figures 1-8, Figure 16, Figure 17). In particular, RIVPA (SEQ ID NO. 5) relaxes the damage caused by radiation (Figures 1, 2) and is systemically active and responsive to chemotherapy (Figures 3, 4, 5 and 6) With significant protective effects observed in the gastrointestinal tract. In addition, IDR is effective in both immunocompetent and leukopenia-deprived animals - suggesting that the GI component of the ARS and the associated hematopoietic effects will not aggravate the efficacy of RIVPA (SEQ ID NO. 5). In view of its anti-infectious role, RIVPA (SEQ ID NO. 5) may also be effective in the hematopoietic sub-symptom of ARS, but its only direct evaluation was the use of intraperitoneal administration of the second-line. RIVPA (SEQ ID NO. 5) has not been predicted to pharmacokinetically interfere with combination therapy and has not been shown to induce major classes of antibiotics (data not shown). RIVPA (SEQ ID NO. 5) does not interfere with recovery from leukopenia (FIG. 15). (SEQ ID NO. 5) and other IDR treatments in combination with the proven safety of RIVPA (SEQ ID NO. 5) in human volunteer studies (see 4.a.iv above) It strongly supports the reduction of acute mortality by use.

Reference

Table 1

                         SEQUENCE LISTING <110> Soligenix, Inc.        Donini, Oreola        Rozek, Annett        Lee, Jackson        North, John        Abrams, Michael   <120> Novel Peptides and Analogs for Use in the Treatment of Oral        Mucositis <130> 502.223WO <140> PCT / US14 / 50516 <141> 2014-08-11 <150> 61/877767 <151> 2013-09-13 <160> 92 <170> PatentIn version 3.5 <210> 1 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Immunological Peptide <400> 1 Lys Ser Arg Ile Val Pro 1 5 <210> 2 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) <223> Xaa is equal to Acetylated K <400> 2 Xaa Ser Arg Ile Val Pro 1 5 <210> 3 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 3 Ser Arg Ile Val Pro Ala 1 5 <210> 4 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 4 Ser Arg Ile Val Ala 1 5 <210> 5 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 5 Arg Ile Val Pro Ala 1 5 <210> 6 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 6 Lys Ile Val Pro Ala 1 5 <210> 7 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE &Lt; 222 > (5) <223> Xaa is equal to D-Amino Acid of Ala <400> 7 Arg Ile Val Pro Xaa 1 5 <210> 8 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 8 Arg Val Pro Ala One <210> 9 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 9 Arg Ile Pro Ala One <210> 10 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE &Lt; 222 > (5) <223> Xaa is equal to Ala-OH <400> 10 Arg Ile Val Pro Xaa 1 5 <210> 11 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 11 Arg Ala Val Pro Ala 1 5 <210> 12 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 12 Arg Arg Ile Val Pro Ala 1 5 <210> 13 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 13 Arg Lys Val Pro Ala 1 5 <210> 14 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 14 Arg Ile Val Pro Lys 1 5 <210> 15 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 15 Arg Pro Val Pro Ala 1 5 <210> 16 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 16 Arg Ile Pro Pro Ala 1 5 <210> 17 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 17 Arg Ile Val Pro Pro 1 5 <210> 18 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 18 Arg Ile Val Pro Gly Gly Ala 1 5 <210> 19 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 19 Gly Gly Ile Val Pro Ala 1 5 <210> 20 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 20 Gly Ile Val Pro Ala 1 5 <210> 21 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 21 Ala Gly Val Ala 1 5 <210> 22 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 22 Arg Ile Val Pro Gly 1 5 <210> 23 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 23 Arg Ile Val Pro Ser 1 5 <210> 24 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 24 Arg Ile Val Pro Leu 1 5 <210> 25 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 25 Arg His Val Ala 1 5 <210> 26 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 26 Arg Ile Pro Val Ala 1 5 <210> 27 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 27 Arg Val Ile Pro Ala 1 5 <210> 28 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 28 Arg Ile Ile Pro Ala 1 5 <210> 29 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 29 Ala Val Pro Ile Arg 1 5 <210> 30 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 30 Ala Pro Val Ile Arg 1 5 <210> 31 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 31 Arg Ile Val Pro Ala 1 5 <210> 32 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) (7) <223> cyclic peptide <400> 32 Cys Arg Ile Val Pro Ala Cys 1 5 <210> 33 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (2) (2) <223> Xaa is equal to I with an NMe backbone <400> 33 Arg Xaa Val Pro Ala 1 5 <210> 34 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE &Lt; 222 > (5) <223> Xaa is equal to A with an N-methylated backbone <400> 34 Arg Ile Val Pro Xaa 1 5 <210> 35 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 35 Arg Ile Val Pro Phe 1 5 <210> 36 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) <223> Xaa is equal to Citrulline <400> 36 Xaa Ile Val Pro Ala 1 5 <210> 37 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 37 Arg Leu Val Pro Ala 1 5 <210> 38 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 38 His Ile Val Pro Ala 1 5 <210> 39 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 39 Ile Arg Arg Val Ala 1 5 <210> 40 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 40 Ala Arg Val Pro Ala 1 5 <210> 41 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 41 Ile Arg Val Pro Ala 1 5 <210> 42 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) <223> Xaa is equal to Ornithine <400> 42 Xaa Ile Val Pro Ala 1 5 <210> 43 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 43 Ser Ile Val Pro Ala 1 5 <210> 44 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 44 Val Ser Ile Ile Lys Pro Ala Arg Val Ser Ser Leu Leu 1 5 10 <210> 45 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 45 Lys Pro Ala Arg Val Pro Ser 1 5 <210> 46 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 46 Arg Val Pro Ser Leu Leu 1 5 <210> 47 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 47 Lys Pro Arg Ala Val Pro 1 5 <210> 48 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 48 Pro Ala Arg Val Pro 1 5 <210> 49 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 49 Ile Arg Val Pro One <210> 50 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 50 Arg Val Pro Ser One <210> 51 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 51 Arg Val Pro One <210> 52 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 52 Pro Ser Val Pro Gly Ser 1 5 <210> 53 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 53 Gly Leu Lys His Pro Ser 1 5 <210> 54 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 54 Arg Ile Val Pro Ala Ile Pro Val Ser Leu Leu 1 5 10 <210> 55 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) Xaa is selected from the group consisting of K, H, R, S, T, O,        Cit, Hci, Dab, Dpr, or glycine based compounds with basic        functional groups substituted on the N-terminal, hSer, Val        (betaOH) <220> <221> MISC_FEATURE <222> (2) (2) &Lt; 223 > Xaa is selected from the group consisting of V, I, K, P, and H <400> 55 Xaa Xaa Pro One <210> 56 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) Xaa is selected from the group consisting of K, H, R, S, T, O,        Cit, Hci, Dab, Dpr, or glycine based compounds with basic        functional groups substituted on the N-terminal, hSer, Val        (betaOH) <220> <221> MISC_FEATURE <222> (2) (2) I, L, V, K, P, G,        H, R, S, O, Dab, Dpr, Cit, Hci, Abu, Nva, Nle and where Xaa can        be N-methylated <220> <221> MISC_FEATURE &Lt; 222 > (3) <223> Xaa is selected from the group consisting of I, V, P <400> 56 Xaa Xaa Xaa Pro One <210> 57 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) P, I, R, C, T, L, and X are selected from the group consisting of S,        V, A, G, K, H, R, O, C, M and F <220> <221> MISC_FEATURE <222> (2) (2) Xaa is selected from the group consisting of K, H, R, S, T, O,        Cit, Hci, Dab, Dpr, or glycine based compounds with basic        functional groups substituted on the N-terminal, hSer, Val        (betaOH) <220> <221> MISC_FEATURE &Lt; 222 > (3) I, L, V, K, P, G,        H, R, S, O, Dab, Dpr, Cit, Hci, Abu, Nva, Nle and where Xaa can        be N-methylated <220> <221> MISC_FEATURE <222> (4) (4) <223> Xaa is selected from the group consisting of I, V, P <400> 57 Xaa Xaa Xaa Xaa Pro 1 5 <210> 58 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) Xaa is selected from the group consisting of K, H, R, S, T, O,        Cit, Hci, Dab, Dpr, or glycine based compounds with basic        functional groups substituted on the N-terminal, hSer, Val        (betaOH) <220> <221> MISC_FEATURE <222> (2) (2) I, L, V, K, P, G,        H, R, S, O, Dab, Dpr, Cit, Hci, Abu, Nva, Nle and where Xaa can        be N-methylated <220> <221> MISC_FEATURE &Lt; 222 > (3) <223> Xaa is selected from the group consisting of I, V, P <220> <221> MISC_FEATURE &Lt; 222 > (5) Xaa is selected from the group consisting of A, A *, G, S, L, F,        K, C, I, V, T, Y, R, H, O, and M,        ofe <400> 58 Xaa Xaa Xaa Pro Xaa 1 5 <210> 59 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) Xaa is selected from the group consisting of K, I, R, H, O, L, V,        A, and G <220> <221> MISC_FEATURE <222> (2) (2) S, P, R, T, H, K, O,        L, V, A, G, S, I <220> <221> MISC_FEATURE &Lt; 222 > (3) Xaa is selected from the group consisting of K, H, R, S, T, O,        Cit, Hci, Dab, Dpr, or glycine based compounds with basic        functional groups substituted on the N-terminal, hSer, Val        (betaOH) <220> <221> MISC_FEATURE <222> (4) (4) I, L, V, K, P, G,        H, R, S, O, Dab, Dpr, Cit, Hci, Abu, Nva, Nle and where Xaa can        be N-methylated <220> <221> MISC_FEATURE &Lt; 222 > (5) <223> Xaa is selected from the group consisting of I, V, P <400> 59 Xaa Xaa Xaa Xaa Xaa Pro 1 5 <210> 60 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) Xaa is selected from the group consisting of S, R, K, H, O, T, I,        L, V, A, and G <220> <221> MISC_FEATURE <222> (2) (2) Xaa is selected from the group consisting of K, H, R, S, T, O,        Cit, Hci, Dab, Dpr, or glycine based compounds with basic        functional groups substituted on the N-terminal, hSer, Val        (betaOH) <220> <221> MISC_FEATURE &Lt; 222 > (3) I, L, V, K, P, G,        H, R, S, O, Dab, Dpr, Cit, Hci, Abu, Nva, Nle and where Xaa can        be N-methylated <220> <221> MISC_FEATURE <222> (4) (4) <223> Xaa is selected from the group consisting of I, V, P <220> <221> MISC_FEATURE <222> (6) V, I, L, G, K, H, X,        R, O, S, T, and F <400> 60 Xaa Xaa Xaa Xaa Pro Xaa 1 5 <210> 61 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 61 Arg Ile Val Pro Ala Cys 1 5 <210> 62 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 62 Arg Arg Val Pro One <210> 63 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE &Lt; 222 > (5) <223> Xaa is equal to A-NHOH <400> 63 Arg Ile Val Pro Xaa 1 5 <210> 64 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 64 Arg Ile Val Pro Ala 1 5 <210> 65 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 65 Arg Ile Gly Pro Ala 1 5 <210> 66 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (4) (4) <223> Xaa is equal to Pip <400> 66 Arg Ile Val Xaa Ala 1 5 <210> 67 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (4) (4) <223> Xaa is equal to Thz <400> 67 Arg Ile Val Xaa Ala 1 5 <210> 68 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (4) (4) <223> Xaa is equal to Fpro <400> 68 Arg Ile Val Xaa Ala 1 5 <210> 69 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (4) (4) <223> Xaa is equal to Dhp <400> 69 Arg Ile Val Xaa Ala 1 5 <210> 70 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 70 Arg Ile His Pro Ala 1 5 <210> 71 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 71 Arg Ile Trp Pro Ala 1 5 <210> 72 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 72 Arg Ile Val Pro Trp 1 5 <210> 73 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 73 Ser Pro Val Ile Arg His 1 5 <210> 74 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 74 Cys Pro Val Ile Arg His 1 5 <210> 75 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 75 Arg Ile Glu Pro Ala 1 5 <210> 76 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 76 Arg Ile Val Pro Glu 1 5 <210> 77 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 77 Arg Ile Val Pro His 1 5 <210> 78 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 78 Arg Ser Val Pro Ala 1 5 <210> 79 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 79 Glu Arg Ile Val Pro Ala Gly 1 5 <210> 80 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 80 Lys Val Ile Pro Ser 1 5 <210> 81 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 81 Lys Val Val Pro Ser 1 5 <210> 82 <211> 4 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 82 Lys Pro Arg Ser One <210> 83 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 83 Arg Ile Pro One <210> 84 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (1) <223> Xaa is equal to Orn <400> 84 Xaa Val Pro One <210> 85 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 85 Ser Val Pro One <210> 86 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 86 Lys Val Pro One <210> 87 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 87 Arg Arg Pro One <210> 88 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 88 Gly Val Pro One <210> 89 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <400> 89 Lys His Pro One <210> 90 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (6) <223> Xaa is equal to D-amino acid Tyr <400> 90 Arg Ile Val Pro Ala Xaa 1 5 <210> 91 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (2) (2) <223> Xaa is equal to tBG, tert-butyl glycine <220> <221> MISC_FEATURE <222> (6) <223> Xaa is equal to tBG, tert-butyl glycine <400> 91 Arg Xaa Val Lys Arg Xaa Val 1 5 <210> 92 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> IMMUNOLOGICAL PEPTIDE <220> <221> MISC_FEATURE <222> (4) (4) Xaa is equal to mp2, 4-Amino-1-methyl-1H-pyrrole-2-carboxylic acid        acid <220> <221> MISC_FEATURE <222> (6) <223> Xaa is equal to NH2 <400> 92 Arg Ile Val Xaa Ala Xaa 1 5

Claims (20)

CLAIMS 1. A method of treating oral mucositis in a subject exposed to radiation or a chemotherapeutic agent in a damaging amount,
a) a peptide comprising the amino acid sequence of amino acids of more than _{7, X 1 X 2 X 3 P} (SEQ ID NO: 56) ( herein, X1 is R a; X2 is I or V, where X2 is X3 is I or V, wherein X3 can be N-methylated; P is a proline or proline analogue; SEQ ID NO: 56 is the first four amino acids of the N-terminus of the peptide ) Or a pharmaceutical salt, ester or amide thereof and a pharmaceutically acceptable carrier, diluent or excipient; or
b) a polynucleotide having the nucleotide sequence of SEQ ID NO: 5, 7, 10, 14, 17, 18, 22, 23, 24, 27, 28, 31, 34, 35, 63, 64, 66-69, 72, 92, or a pharmaceutical salt, ester or amide thereof, and a pharmaceutically acceptable carrier, diluent or excipient. The method of claim 1, wherein the peptide is SEQ ID NO: 5 or a pharmaceutical salt, ester or amide thereof and a pharmaceutically acceptable carrier, diluent or excipient. 3. The method of claim 1, wherein the peptide is administered by oral, subcutaneous, intramuscular, intravenous, transdermal, intranasal, pulmonary administration, or osmotic pumps. 2. The method of claim 1, wherein the effective amount of the peptide is at least 1.5 mg / kg. 2. The method of claim 1, wherein the peptide is administered to the patient every three days during the administration of radiation or chemotherapeutic agents. 3. The method of claim 1, wherein the peptide is administered to the patient in combination with a locally active corticosteroid or metabolite thereof in oral dosage form, wherein the oral dosage form is effective for local or local treatment of the gastrointestinal tract and oral cavity of the patient, Wherein the patient further exhibits symptoms of inflammation due to tissue damage resulting from radiation or chemotherapy treatment. 7. The method of claim 6 wherein the locally active corticosteroid is beclomethasone dipropionate. 7. The method of claim 6, wherein the metabolite is 17-beclomethasone monopropionate. 7. The method of claim 6, wherein the effective amount of locally active corticosteroid is 8 mg / day. 2. The method of claim 1, wherein the peptide is administered in combination with an effective amount of an antibiotic. Amino acid SEQ ID NO. 91 or SEQ ID NO. 92 or a pharmaceutical salt, ester or amide thereof. Wherein tBg is tertiary-butylglycine, and further wherein R (tBg) V2 is an amino acid sequence of the side chain lysine amino group and the amino acid sequence of SEQ ID NO: RTI ID = 0.0 &gt; V2. &Lt; / RTI &gt; Amino-1-methyl-lH-pyrrole-2-carboxylic acid (SEQ ID NO: 92)

&Lt; / RTI &gt; A method of treating an individual suffering from a disease selected from the group consisting of bacterial infections, mucositis and colitis, or exposure to acute radiation, including administering to the individual a peptide consisting of the amino acid sequence of Table 1. 15. The method of claim 14, wherein the amino acid sequence is SEQ ID NO. 5, SEQ ID NO. 91 and SEQ ID NO. 92. &Lt; / RTI &gt; 16. The method of claim 15, wherein the disease is a bacterial infection. 16. The method of claim 15, wherein the amino acid sequence is SEQ ID NO. 91 or SEQ ID NO. 92, and wherein the peptide is administered in combination with an effective amount of an antibiotic. A method for preventing infection in a subject,
Effective amount of
a) a peptide comprising the amino acid sequence of amino acids less than _{7, X 1 X 2 X 3 P} (SEQ ID NO: 56) ( herein, X1 is R a; X2 is I or V, where X2 is X3 is I or V, wherein X3 can be N-methylated; P is a proline or proline analogue; SEQ ID NO: 56 is the first four amino acids of the N-terminus of the peptide ) Or a pharmaceutical salt, ester or amide thereof and a pharmaceutically acceptable carrier, diluent or excipient; or
b) a polynucleotide having the nucleotide sequence of SEQ ID NO: 5, 7, 10, 14, 17, 18, 22, 23, 24, 27, 28, 31, 34, 35, 63, 64, 66-69, 72, 92, or a pharmaceutical salt, ester or amide thereof, and a pharmaceutically acceptable carrier, diluent or excipient. 16. The method of claim 15, wherein the peptide is selected from the group consisting of SEQ ID NO. 91 or SEQ ID NO. RTI ID = 0.0 &gt; 92. &Lt; / RTI &gt; 20. The method of claim 19, wherein the peptide is administered in combination with an effective amount of an antibiotic.

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